A landmark study comparing 2D and 3D models for DILI prediction
In their peer-reviewed paper, titled “Utility of spherical human liver microtissues for prediction of clinical drug-induced liver injury”, Will Proctor, Alison Foster, Simon Messner, Dominic Williams, and several other notable experts in the field, published a landmark study comparing 2D and 3D DILI models, highlighting the advantages of using 3D InSight™ Human Liver Microtissues (3D hLiMT) for DILI prediction.
The scientific paper is a highly recommended resource for anyone in the field of Liver Safety. Here is why:
Publication Objective and Study Design
The authors set out to perform a systematic validation of 3D hLiMT vs. 2D PHH for their predictive value in discriminating between known hepatotoxicants and clinically safe drugs. They tested a panel of 110 clinically known drugs on 2D Primary Human Hepatocytes (PHH) and on InSphero's human-derived liver spheroids, utilizing the same hepatocyte lot, compound concentrations, and endpoint (ATP-content).
Key Findings about 3D InSight™ Human Liver Microtissues for DILI Prediction
The authors found that the 3D InSight™ liver spheroids were twice as sensitive in identifying known hepatotoxicants as compared to 2D PHH. The specificity for prediction of non-DILI drugs also remained quite high (90%), even after 14 days of compound exposure. Furthermore, the model enabled the assessment of novel hepatic injury biomarkers, such as miRNA122, HMGB1, and α-GST.
Figure 1: DILI Prediction with 3D InSight™ Liver Microtissues - Graph showing a 2-fold higher sensitivity of liver spheroids in predicting DILI-positive compounds.
About the Data
In their paper comparing 2D and 3D models for DILI prediction, AstraZeneca and Genentech chose the validation compound set to represent a “real-world” situation. To conduct a truly independent assessment of the model, all 3D studies were performed at InSphero under conditions where the identities of the 110 compounds were blinded, and all 2D studies were conducted independently by Genentech.
The compound set included some with limited solubility, as well as many weak DILI compounds with known rare and late DILI onset. For example, out of the 21 DILI compounds where no IC50 value was determined, 14 compounds were only tested at less than 20-fold of the patients’ plasma concentrations due to solubility issues, thus preventing calculations of margin-of-safety.
Four drugs (Stavudine, Methotrexate, Dantrolene, Nifedipine) were toxic in vivo only after several months of administration, and another drug (Trovafloxacin) is known to cause DILI in an idiosyncratic reaction, i.e., in the presence of inflammation, thus requiring different stimuli. Trovafloxacin toxicity was, however, correctly detected in the presence of an inflammatory stimulus in a separate study. Read more>>
About the Model: 3D InSight™ Human Liver Microtissues for DILI prediction
Our 3D InSight™ Human Liver Microtissues have several features that make them a superior choice for toxicity testing:
- Polarized hepatocytes with active hepatobiliary bile-salt secretion (BSEP and MRP2) and the presence of bile canaliculi
- Preserved cytochrome P450 activity over the 14-day assay period
- 6-fold increase in mitochondrial spare respiratory capacity compared to 2D PHH
- Incorporation of inflammation-sensing Kupffer cells for detection of indirect effects on liver function
Viable for more than 28 days in culture and with more physiologically relevant biology than 2D PHH, the human liver spheroids enable cumulative assessment of the most important factors driving DILI in vivo:
- Long-term drug exposure
- Active drug metabolism
- BSEP inhibition
- Mitochondrial impairment
Our 3D InSight™ Human Liver Microtissues are delivered fully QC’d and assay-ready in an automation-compatible 96-well format to ensure seamless integration into your laboratory workflows. The plate features our unique SureXchange™ ledge that eliminates spheroid loss during media exchange and drug dosing. This cost-effective and efficient solution for predictive DILI testing allows maximal confidence in decision-making and efficiency in drug safety evaluation.
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